Los autores agradecen a Laurie von Melchner, Hospital Universitario de Berna, Departamento de Neurocirugía, Berna, Suiza, para la revisión y edición del manuscrito y Paskus Jeremías, Hospital de Niños de Boston, Boston, MA corrección de pruebas para el proyecto inicial. Apreciamos el hábil manejo de cuidado de los animales, la anestesia, y la asistencia operativa de Daniel Mettler, DVM, Max Müller, DVM, Daniel Zalokar y Olgica Beslac, Instituto de Cirugía Experimental, Departamento de Investigación Clínica de la Universidad de Berna, Berna, Suiza. Damos las gracias a Michael Lensch, Jefe de Investigación Enfermera del Departamento de Medicina de Cuidados Intensivos del Hospital Universitario de Berna y de la Universidad de Berna, Berna, Suiza, para el monitoreo de datos en tiempo real y post-procesamiento de los parámetros fisiológicos. Agradecemos a Edin Nevzati, Carl Muroi, y Salomé Erhardt, por su excelente asistencia técnica y operativa de laboratorio. Este trabajo fue apoyado por el Departamento de IntensivCuidado de Medicina e, Hospital de la Universidad de Berna y la Universidad de Berna, Berna, Suiza, el Departamento de Investigación Clínica de la Universidad de Berna, Berna, Suiza, y el Fondo de Investigación del Hospital Cantonal de Aarau, Aarau, Suiza. Agradecemos a Elsevier, para pedir el permiso de las figuras 1 y 2.

Tres meses de edad hembra conejos de Nueva Zelanda que pesan 2.5 a 3.5 kg se utilizaron para este procedimiento. El estudio se realizó de acuerdo con los Institutos Nacionales de Salud directrices para el cuidado y uso de animales de experimentación y con la aprobación del Comité de Cuidado de Animales del cantón de Berna, Suiza (aprobación N. ° 105/13). Todos los procedimientos quirúrgicos se realizaron bajo condiciones estériles en el Instituto de Cirugía Experimental del Departamento de Investigación Clínica del Hospital Universitario de Berna en Berna, Suiza. Un anestesiólogo veterinario supervisa los animales durante la cirugía y durante la recuperación. 1. Preparación de los animales, Posicionamiento y Arteria subclavia Canulación <ol><li> Inducir la anestesia general en el conejo con la inyección intramuscular de ketamina (30 mg / kg; Ketalar, 50 mg / ml) y xilazina (6 mg / kg; Xylapan 20 mg / ml) y la profundidad de control de la anestesia mediante la comprobación de la respuesta del conejo para nocivo stimulatión (por ejemplo, una pizca dedo del pie). Ver 1.7 en caso de respuesta positiva. </li><li> Tire hacia abajo el párpado inferior de ambos ojos y aplique una pequeña cantidad de ungüento en los párpados para evitar la sequedad y la irritación adicional. </li><li> Cateterizar la vena lateral de la oreja con un Gbutterfly intravenoso periférico 20 (20 catéter vascular G), fijar con cinta adhesiva, y conectarse a una bolsa de gravedad que contiene cloruro de sodio al 0,9% (500 ml / 24 hr) y ketamina (40 mg / kg / hr) / xilazina (4 mg / kg / h) para la administración intravenosa continua (iv) la anestesia. Administrar analgésicos adicional cada 15 min iv (fentanilo, 1 mcg / kg). Nota: Evite los anestésicos volátiles de gas, que se asocia con la reducción de la PPE, el aumento de la CBF, y la disminución de la tasa metabólica cerebral de oxígeno 11 anestésicos intravenosos proporcionan características más idóneas para neuroanestesia preservando CBF y la vasoconstricción cerebral, 12, que tiene una importancia superior al estudiar cerebral. vasoespasmo. Además, aunque la mortalidad se incrementa en SPONTanimales respirar aneously, puede imitar mejor la situación humana de la HSA aguda. </li><li> Proporcionar oxígeno (1 - 2 L / min) a través de una máscara respiratoria que permite dióxido de carbono al final de las mareas (ETCO 2) seguimiento. </li><li> Instale un electrocardiograma de 3 canales (ECG) .place tres electrodos subcutáneos en una disposición triangular en la parte ventral del conejo; específicamente, colocar un electrodo sobre la región centrotorácico derecha (con la distancia al campo afeitado estéril para la canulación de la arteria subclavia) y dos electrodos en el abdomen inferior distribuidos en ambas extremidades. </li><li> Monitorear la profundidad de la anestesia cada 15 minutos durante la cirugía siguiendo la frecuencia respiratoria, frecuencia cardíaca (FC) monitoreado desde la señal de ECG, y la reacción a la estimulación nociva. </li><li> En caso de respuesta positiva a la estimulación nociva (pizca dedo del pie), adaptar la profundidad de la anestesia mediante bolo de ketamina (6 mg / kg) iv en bolo y xilazina (0,05 mg / kg) iv y / o un bolo analgesia adicional con Fentanilo (1 mcg / kg) iv </li><li> Fijar el conejo en una posición supina sobre una placa de calentamiento cuerpo, inclinando la cabeza 20 ° hacia abajo y girando ligeramente contralateral al lado en el que se expone la arteria subclavia. </li><li> aplicar pomada para los ojos y preparar la zona para la cirugía por el afeitado el pelo sobre el músculo pectoral derecho alrededor del tercio medio de la clavícula, y sobre el cráneo frontal-, parietal- y occipital, el cuello, y sobre la arteria femoral común derecha. </li><li> Desinfectar la piel durante 3 min con un amplio espectro antiséptico, por ejemplo., Povidona-yodo. </li><li> Cubrir el conejo con sábanas estériles. Realizar todos los procedimientos adicionales en condiciones estériles y con frecuencia aplicar 4% papaverina HCl y solución de antibiótico (sulfato de neomicina 5 mg / ml) por vía tópica para prevenir el vasoespasmo arterial por la manipulación del vaso y las infecciones locales. </li><li> Infiltrarse en el músculo pectoral con anestésicos locales (lidocaína 1% de la máxima 6 mg / kg). Hacer una incisión en la piel paraesternal y preparar elmúsculo pectoral. Usando el microscopio, diseccionar la arteria subclavia y seguro con una ligadura proximal y distal (4-0 suturas multifilamento) alrededor del extremo expuesto. Mantenga una ligadura cerca del control proximal en el lugar para asegurar el catéter y ligar el vaso distal. </li><li> Realizar una arteriotomía en la pared de la arteria subclavia por la incisión de la arteria con un microscissor curvada y canular la arteria subclavia retrógradamente con un pequeño intravasal de 3 vías llave de paso. Asegurar el catéter por ligadura doble nudo hacia la ligadura distal con el fin de impedir la torsión arterial o la flexión de la parte proximal de la arteria y para evitar el sangrado deslizamiento o masiva. </li></ol> 2. la presión arterial y Monitoreo La gasometría arterial <ol><li> Conecte el stop de 3 vías polla para i) la cánula intravasal de gases en sangre arterial (ABG) analiza, pH, PaCO 2, PaO 2, bicarbonato, exceso de base, y SO 2, ii) la sangre arterial invasivadispositivo de medición de presión, y iii) el dispositivo de derivación. </li><li> Recoger muestras de sangre para el estado de ABG (PaCO 2, PaO 2) y un seguimiento continuo de los parámetros estándar cardiovasculares y respiratorias (presión arterial, de recursos humanos, ECG, frecuencia respiratoria y telerrespiratorias CO 2) y de transferencia de datos a través de la interfaz de salida analógica a un analógico- convertidor A / registrador de datos digital y tienda. NOTA: Las presiones se centraron en los niveles del corazón antes y después de cada sesión, y calibración de la presión de la analógica / digital convertidor y el sistema de registro de datos se llevará a cabo una vez antes de que comience la serie. </li></ol> 3. Base angiografía por sustracción digital <ol><li> Coloque un dispositivo de encolado externo (pequeña esfera) sobre ambos ángulos de la mandíbula con el fin de calibrar el angiograma. NOTA: Esto permite la comparación exacta de las mediciones post hoc de la línea de base y seguimiento diámetro del vaso. </li><li> Realizar la angiografía por sustracción digital (DSA) por retrógrada dentro de uninyección en bolo rterial de Iopamidol no iónico (0,6 ml / kg, 5 ml / seg durante 2 segundos) a través de la arteria canulada y enjuagar la cánula inmediatamente después de la inyección en bolo con solución salina con el fin de evitar la oclusión de este último. </li><li> Obtener imágenes (7 imágenes en 14 segundos) del sistema vertebrobasilar utilizando una rápida grabación angiografía secuencial en una posición oblicua 5 ° izquierda-anterior. </li><li> Infíltrate en la zona alrededor de la arteria femoral común derecha con anestésicos locales (lidocaína al 1%, máximo de 6 mg / kg). Haga una pequeña incisión en la piel inguinal. Usando el microscopio para la visualización, diseccionar la arteria femoral común y seguro con una ligadura proximal y distal (4-0 suturas multifilamento) alrededor del extremo expuesto. </li><li> Después de arteriotomía, canular la arteria femoral con una vaina de 5-F. Enjuague el puerto lateral de la funda con solución salina. </li><li> Avanza un catéter 5-F en la arteria innominada a través de la vaina bajo fluoroscopia. Crear una hoja de ruta, para luego avanzar un alambre de guía a thsistema vertebrobasilar e. Inyectar un bolo de Iopamidol no iónico (0,6 ml / kg, 5 ml / seg en 2 seg) para DSA como se describe en el paso 3.2. </li></ol> 4. rotación a la posición propensa <ol><li> Siguiendo la línea de base DSA, vuelva a colocar el conejo de posición supina a la posición prona. Tenga cuidado de no manipular o cambiar de lugar la posición de los catéteres intra-arteriales. </li><li> Coloque la cabeza en un soporte del cabezal en un ángulo de 30 °, la cabeza orientada hacia abajo. </li></ol> 5. Cisterna Magna Punción <ol><li> Desinfectar la piel que cubre la cabeza y el cuello con povidona yodada 3 veces durante 1 minuto cada uno, y cubrir el área quirúrgica con sábanas estériles. </li><li> Insertar un 22 G x 40 mm aguja de acceso espinal infantil transcutánea en la cisterna magna sin ninguna incisión en la piel antes o desplazamiento del músculo. </li><li> Confirme que el animal esté totalmente anestesiado, garantizando una falta de respuesta del dedo del pie-pellizco antes de deslizar la aguja hacia abajo a lo largo del hueso protuberancia occipital externahasta que se detecta una brecha; no empuje la aguja más. </li><li> Confirmar el correcto posicionamiento de la aguja mediante la observación de goteo espontáneo de líquido cefalorraquídeo con la cabeza del conejo inclinado hacia abajo en un ángulo de 20 ° para unos pocos min. </li></ol> 6. Instalación de la presión intracraneal y Monitoreo del flujo sanguíneo cerebral <ol><li> Después de la piel en la línea media y la incisión galea, insertar un pequeño retractor quirúrgico. </li><li> Hacer tres osteotomías ronda (2 mm de diámetro) utilizando un microtaladro de alta velocidad en la parte frontal del cráneo de acuerdo con los puntos de referencia del cráneo exteriores (Figura 1) 9, es decir, sobre el bulbo olfativo y frontal bilateral para la colocación de un dispositivo de neuromonitorización si necesario. Use una regla escala milimétrica para determinar las coordenadas de rebabas colocación agujero de la siguiente manera: monitorización de la presión intracraneal (PIC) en la línea midpupillary, una o dos mm de la línea sagital medio; sondas láser-Doppler intraparenquimatosascuatro y cincuenta y seis mm anterior y lateral a la bregma (Figura 1). </li><li> Visualice la duramadre, y realizar una hemostasia meticulosa: utilizar cera de hueso para la hemostasis ósea en virtud de su acción taponamiento y realizar la hemostasia local mediante coagulación bipolar de la duramadre. </li><li> Coloque la presión intracraneal (ICP) de punta monitor de intraparenquimatosa en el bulbo olfatorio derecho a una profundidad de 2 mm y luego calibrar. </li><li> Coloque las sondas con aguja fina los dos laser-Doppler flujometría utilizando un sistema de abrazadera externa e insertarlos en los agujeros de trépano correspondientes, tanto en los hemisferios derecho e izquierdo frontales lateral al sistema ventricular, es decir, en la línea media para evitar interferencias con el líquido cefalorraquídeo. Coloque sondas de aguja a una profundidad de 2,5 mm. </li><li> Después de la colocación de las sondas de neuromonitorización, selle todos los orificios de trepanación con un tapón grueso de cera de hueso para mantener el cráneo estanca a los fluidos. </li><li> Medir los parámetros basales de la presión arterial media (MAP), ICP y el flujo sanguíneo cerebral (CBF) usando un monitor multiparamétrico y de cuatro canales tejido láser-Doppler monitor de perfusión de la sangre. </li></ol> 7. Derivación Inducción <ol><li> Conecte la aguja de acceso espinal en la cisterna magna de la arteria subclavia previamente sondaje mediante un tubo de control de presión llena de sangre. Utilice la llave de paso de 3 vías para la medición de la presión arterial y como puerto de muestreo de sangre. NOTA: La gravedad de SAH depende de la cantidad de sangre, y se puede estimar más o menos por la extensión de la formación de coágulos subaracnoideos en el momento de la cosecha cerebro 5,11. </li><li> Monitorear continuamente PAM, FC, ECG, frecuencia respiratoria y la final de la espiración de CO 2 a una velocidad de muestreo de 1 Hz de 6 minutos antes hasta por lo menos 20 minutos después de la HSA. </li><li> Confirmar que el animal está totalmente anestesiado asegurando una falta de respuesta pizca dedo del pie antes de abrir la conexión de derivación entre la arteria subclavia y la cisterna magna para inducir SAH por la presión gradient. NOTA: A HSA controlado puede lograrse mediante el cierre de la derivación en cualquier punto de tiempo (por ejemplo, a un nivel deseado de ICP.). </li><li> Registre los valores en estado estacionario durante un período de tiempo de aproximadamente 15 min. </li><li> Después de ICP alcanza su punto máximo, mantenga la aguja de acceso espinal en su lugar hasta que la PIC vuelve a un estado estacionario cerca de los valores basales. Si se mantiene la meseta ICP durante más de 10 segundos o si la PIC disminuye de forma espontánea, cierra la derivación. </li><li> Retire CBF sondas con aguja fina y la sonda ICP, tapar los agujeros de trépano con cera de hueso, quitar todos los catéteres (incluyendo catéter subclavia, desde la manipulación del catéter con sangrado consecutivo se asocia con una alta morbilidad y mortalidad, y aumenta la tasa de infección), lleve a cabo el lavado de heridas rigurosa con El sulfato de neomicina y suturar la piel. </li></ol> 8. Tratamiento postoperatorio <ol><li> El procedimiento tiene una duración de aproximadamente 2 horas. Debido a la vida media de ketamina y xilazina, el tiempo de recuperación del ánimal es bastante corto - aproximadamente 1 hr. Los animales se mantuvieron bajo una lámpara de calentamiento durante la recuperación. Líquidos adicionales no se proporcionan. Durante esta fase inicial de la recuperación postoperatoria, aplicar buprenorfina 0,02 mg / kg sc cada 8 horas durante 24 horas. </li><li> Aplicar parches de matriz de fentanilo transdérmico de liberación de 12,5 mg / hr en la región del cuello afeitada de los animales para la analgesia efectiva durante el próximo 72 hr. </li><li> No deje un animal sin vigilancia hasta que se haya recuperado el conocimiento suficiente para mantener decúbito esternal. </li><li> No devuelva un animal que ha sido sometido a cirugía en la compañía de otros animales hasta que se recupere completamente. </li></ol> 9. Seguimiento angiografía por sustracción digital para evaluar DCVS en el Día 3 <ol><li> Realice los pasos 1.1 hasta 3.6 como se describe anteriormente. </li><li> La eutanasia a los animales mediante la inyección intraarterial bolo de thiopenthal de sodio (40 mg / kg) (pentotal, Ospedalia AG, Hünenberg, Suiza). En los casos en que la histología y immunohistochemistry es necesario, realizar una perfusión-fijación intracardíaca a temperatura ambiente a una presión de perfusión de 100 cm H 2 O. </li></ol>

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Genetics. 6, 507-512 (2005).</li><li>Flisikowska, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+immunoglobulin+gene+disruption+and+targeted+replacement+in+rabbit+using+zinc+finger+nucleases.">Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.</a> PloS one. 6, e21045 (2011).</li><li>Nakajima, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+aging+on+cerebral+vasospasm+after+subarachnoid+hemorrhage+in+rabbits.">Effects of aging on cerebral vasospasm after subarachnoid hemorrhage in rabbits.</a> Stroke. 32, 620-628 (2001).</li></ol>

None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

El modelo de shunt produce patología similar a la observada en seres humanos después de la HSA aguda 3,8,10. Se ha sugerido que puede exacerbar EBI, mantener e incluso desencadenar DCVS 12, y como tal, este modelo puede ayudar en la investigación de las fases tempranas y tardías DCVS, incluyendo EBI y DCVS interacciones siguientes SAH. En particular, repetible en vivo DCVS técnicas de monitoreo incluyendo DSA 13, angiografía por tomografía computarizada 14 y Do...

La hemorragia subaracnoidea (HSA) es una de las más potencialmente mortal condiciones neuropatológicos, con frecuencia conduce a un daño neurológico permanente o la muerte 1. Las investigaciones anteriores se ha centrado en el vasoespasmo cerebral retardada (DCVS) como la etiología primaria de los déficits neurológicos asociados con SAH 2. Sin embargo, los pobres en general, los resultados clínicos de los pacientes que sufren de la HSA después del tratamiento del vasoespasmo ha llevado a una expansión del enfoque de la investigación para incluir los efectos de la lesión cerebral temprano (EBI) después de la HSA 3. Una mayor comprensión de la importancia de ambos EBI y DCVS en contribuir a pobres resultados clínicos después de la HSA es esencial para el desarrollo de estrategias terapéuticas más eficaces. Hasta ahora, la inyección de sangre autóloga simple y doble en la cisterna magna ha sido el método estándar para SAH inducción para el estudio de DCVS 2-6. Aunque se utiliza comúnmente en estudios anteriores,este modelo muy probablemente no reproduce los principales cambios neuropatológicos asociados con HSA inducida EBI 7. En contraste, la perforación endovascular es conocida por producir graves cambios fisiopatológicos agudos que imitan parcialmente los síntomas de EBI 7. Este informe describe un nuevo modelo de conejo de SAH diseñada para permitir la investigación de ambos EBI y DCVS, permitiendo así la caracterización más precisa de la patología inducida por SAH 8-10. Con la técnica descrita, el modelo estándar cisterna magna se adapta mediante la conexión del sistema arterial de la arteria subclavia y la cisterna magna a través de una derivación extracorpórea. El flujo de sangre está por lo tanto ligado a la fisiología del conejo y accionado por el gradiente de presión entre la sangre arterial y la presión intracraneal. El sangrado se detiene cuando la presión intracerebral (ICP) es igual a la presión arterial diastólica y la sangre en el sistema de derivación se coagula. Utilizando el host &amp; #8217; s fisiología reduce examinador dependiente de la inducción de la HSA, dando lugar a un modelo más consistente de la HSA que produce de forma fiable tanto EBI y fenotipos DCVS 3,8-10.

<table border="0" cellpadding="0" cellspacing="0" width="905">
	<tbody>
		<tr height="34">
			<td height="34" style="height:35px;width:279px;">Name</td>
			<td style="width:236px;">Company</td>
			<td style="width:223px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Equipment</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">operation microscope</td>
			<td style="width:236px;">Zeiss, Jena, Germany</td>
			<td colspan="2">Zeiss, OPMI-MD surgical microscope</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">surgical equipment</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:223px;">forceps medical n&#176;5, vessel sciccors 8cm, microclip 4mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">respirator</td>
			<td style="width:236px;">Hugo Sachs</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hair clipper</td>
			<td style="width:236px;">3M Surgical Clipper&#160;</td>
			<td style="width:223px;">&#160;Starter Kit 9667A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">body warm plate</td>
			<td style="width:236px;">FHC</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">blood gas analyzer</td>
			<td style="width:236px;">Radiometer, Copenhagen, Denmark</td>
			<td style="width:223px;">ABL 725</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:279px;">cardiac monitoring</td>
			<td style="width:236px;">Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td>
			<td style="width:223px;">AP-05</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">BIOPAC Systems, Inc., Goleta, CA, USA</td>
			<td style="width:223px;">Biopac MP100 and acqKnowledge software,version 3.8.1</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td>
			<td style="width:223px;">image-Pro Plus version&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">angiography apparatus</td>
			<td style="width:236px;">DFP 2000 A-Toshiba</td>
			<td style="width:223px;">MIIXR0001EAA</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ICP monitor</td>
			<td style="width:236px;">Camino Laboratories, San Diego, CA, USA</td>
			<td style="width:223px;">ICP monitor, Model 110-4B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blood flow monitor</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">CAL KIT microsphere solution</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">laser-Doppler flowmetry fine needle probes&#160;</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">MNP110XP, 0.48 mm diameter</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pressure tube</td>
			<td>B. Braun, Germay</td>
			<td style="width:223px;">PE 1.0 mm &#215; 2.0 mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">anesthesia monitor</td>
			<td>GE Medical Systems, Switzerland&#160;</td>
			<td style="width:223px;">Datex S5 Monitor</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Material</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20 G vascular catheter</td>
			<td style="width:236px;">Smiths Medical</td>
			<td style="width:223px;">Jelco i.v. catheter, REF 4057</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">5.5F three-lumen central venous catheter&#160;</td>
			<td style="width:236px;">Connectors, Tagelswangen, Switzerland</td>
			<td>silicone catheter STH-C040</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">22Gx40mm needle&#160;</td>
			<td style="width:236px;">Emergo Group Inc., Netherlands</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-speed microdrill</td>
			<td style="width:236px;">Stryker, Solothurn, Switzerland</td>
			<td>5400-15&#160;</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">bone wax</td>
			<td style="width:236px;">Ethicon, Johnson &#38; Johnson,NJ, USA</td>
			<td style="width:223px;">ETHW31G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">bipolar forceps</td>
			<td style="width:236px;">Aesculap, Inc., PA, US</td>
			<td>US349SP&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Ketamin</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Buprenorphine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fentanyl</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">transdermal fentanyl matrix patches&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lidocaine 1%&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4% papaverin HCl&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neomycin sulfate&#160;</td>
			<td style="width:236px;">Research Organics Inc., OH, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Povidone-iodine&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">0.9% sodium chloride</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Iopamidol&#160;</td>
			<td style="width:236px;">Abott Laboratories, IL, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">3-0 resorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>VCP824G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">4-0 polyfilament sutures</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:223px;">VCP284G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

Lesión cerebral temprana y tardía vasoespasmo cerebral tanto contribuyen a resultados desfavorables después de una hemorragia subaracnoidea (HSA). Modelos animales reproducibles y controlables que simulan ambas condiciones son actualmente poco común. Por lo tanto, se necesitan nuevos modelos con el fin de imitar las condiciones fisiopatológicas humanos como resultado de la HSA. Este informe describe los matices técnicos de un modelo SAH sangre-shunt conejo que permite el control de la presión intracerebral (ICP). Una derivación extracorpórea se coloca entre el sistema arterial y el espacio subaracnoideo, que permite SAH-examinador independiente en un cráneo cerrado. Paso a paso las instrucciones de procedimiento y el equipo necesario se describen, así como las consideraciones técnicas para producir el modelo con un mínimo de mortalidad y morbilidad. Se describen los detalles importantes que se requieren para la creación quirúrgica exitosa de esta robusta, simple y consistente modelo de conejo SAH ICP-controlado.

<html> El modelo de derivación de sangre de conejo de SAH se describe en este informe produce EBI en el hipocampo (Figura 2A, B), la corteza basal (Figura 2A, B), y la vasculatura cerebral (Figura 2C) tan pronto como 24 horas después de la lesión y muestra una característica distribución de la sangre (Figura 2D) 8. Además, el modelo desencadena moderada a severa grados de DCVS en tres días después de la HSA de inducción (Figur...

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

El Conejo Modelo Blood-shunt para el Estudio de la aguda y secuelas tardías de Hemorragia subaracnoidea: Aspectos Técnicos

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

the-rabbit-blood-shunt-model-for-study-acute-late-sequelae

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JoVE Journal

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13.5K vistas.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Video: The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

Microcirugía venosa bolsa arterial-Bifurcación Los aneurismas en el modelo de conejo: Aspectos Técnicos

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

Investigación

Medicina

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarcadores en un modelo animal para Revelar correlatos neurales, hematológicas y conductuales de estrés postraumático

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures3. The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism4-6. This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model7, 8. They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode8. A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours8.
When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)9-11, a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration12 and provides a measure of mitochondrial redox state13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions14, 15 and to monitor the progression of hypoxic conditions over time10.
The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution16 at 37 &deg;C. Aortic, LV, pulmonary artery, and left &amp; right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350&plusmn;25 nm) and used to illuminate the epicardium. Emitted light was filtered (460&plusmn;20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions.

The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

NADH fluorescencia de imágenes de biventricular aislada Trabajo corazones de conejo

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited ...

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

Una baja mortalidad modelo de rata para evaluar vasoespasmo cerebral retardado tras la hemorragia subaracnoidea experimental

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

Humanos neuroendocrinos líneas de células tumorales como un modelo tridimensional para el estudio de la terapia humana tumor neuroendocrino

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Un modelo murino de la implantación de un stent en la arteria carótida para el Estudio de la reestenosis

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspectos técnicos de la aortocava Fístula Ratón

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Un modelo murino de hemorragia subaracnoidea

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

Un modelo murino de restenosis arterial: Aspectos Técnicos del femoral Lesiones alambre

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Medición de la contractilidad isométrica de la arteria mesentérica de ratón con cable miografía

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...